Lactiplantibacillus plantarum monolayer enhanced bactericidal action of carvacrol: biofilm inhibition of viable foodborne pathogens and spoilage microorganisms

Valeria Poscente; Luciana Di Gregorio; Manuela Costanzo; Chiara Nobili; Roberta Bernini; Luigi Garavaglia; Annamaria Bevivino

doi:10.3389/fmicb.2023.1296608

Lactiplantibacillus plantarum monolayer enhanced bactericidal action of carvacrol: biofilm inhibition of viable foodborne pathogens and spoilage microorganisms

Front Microbiol. 2023 Nov 22:14:1296608. doi: 10.3389/fmicb.2023.1296608. eCollection 2023.

Authors

Valeria Poscente^{1

2}, Luciana Di Gregorio¹, Manuela Costanzo¹, Chiara Nobili¹, Roberta Bernini², Luigi Garavaglia³, Annamaria Bevivino¹

Affiliations

¹ Department for Sustainability, Biotechnologies and Agroindustry Division, Italian National Agency for New Technologies, Energy and Sustainable Economic Development (ENEA), Rome, Italy.
² Department of Agriculture and Forest Sciences, University of Tuscia, Viterbo, Italy.
³ I.L.P.A. Group, Valsamoggia, Italy.

Abstract

The prevalence of biofilm-associated microorganisms and the increasing use of ready-to-eat fresh products represent the current duality the food industry must address. Innovative and eco-friendly antibiofilm solutions and appropriate microbiological food control systems are urgently needed to improve food quality and safety. This study aimed to investigate the in vitro combined efficacy of carvacrol with a pre-formed biofilm monolayer of the probiotic Lactiplantibacillus plantarum DSM 20174. The antimicrobial activity of carvacrol against both planktonic and sessile cells of foodborne pathogens and spoilage microorganisms, alone or in the presence of the pre-formed biofilm of L. plantarum, was investigated by culture-based methods along with flow cytometry (FCM) to monitor cells' cultivability and viability. The synergistic action of carvacrol and the pre-formed biofilm of L. plantarum was evaluated in the 96-well plates. The results showed that L. plantarum pre-formed biofilm monolayer enhanced the antimicrobial effect of carvacrol determining a bactericidal action while the treatment alone induced the viable but not culturable (VBNC) cell state only. Furthermore, the great efficacy of the combined treatment allowed the application of a lower concentration of carvacrol (100 ppm) to achieve significant damage in cell viability. In conclusion, the incorporation of carvacrol into the L. plantarum pre-formed biofilm represents a promising alternative for an antimicrobial functionalized ready-to-eat packaging.

Keywords: biofilm control; carvacrol; flow cytometry; food packaging; food spoilage; foodborne pathogens; pre-formed biofilm; probiotics.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was supported by ILIP S.r.l and funded by the European Commission—NextGenerationEU programme under the National Recovery and Resilience Plan (NRRP), Mission 4 “Education and Research,” Component 2 “From research to business:” Project ON Foods—Research and innovation network on food and nutrition Sustainability, Safety and Security—Working ON Foods, Project code PE00000003, CUP I83C22001790001, Investment 1.3 creation of “Partnerships extended to universities, research centers, companies for the financing of basic research projects,” and Project SUS-MIRRI.IT “Strengthening the MIRRI Italian Research Infrastructure for Sustainable Bioscience and Bioeconomy,” Project code IR0000005, CUP D13C22001390001, Investment 3.1 Fund for the realisation of an integrated system of research and innovation infrastructures, Action 3.1.1 “Creation of new research infrastructures strengthening of existing ones and their networking for Scientific Excellence under Horizon Europe.”